scholarly journals USE OF SUCROSE AND MANNITOL FOR DIFFERENTIATION OF FLAX GENOTYPES BY RESISTANCE TO OSMOTIC STRESS

2020 ◽  
Vol 15 (3) ◽  
pp. 10-15
Author(s):  
Elena Vinogradova
Keyword(s):  
Osmotic Stress ◽  
Sucrose Solution ◽  
Primary Root ◽  
Callus Cultures ◽  
Immature Embryos ◽  
Selective Agent ◽  
Scientific Center ◽  
Morphogenic Callus ◽  
Callus Tissues

The studies were carried out with the aim of studying the effect of various concentrations of sucrose and mannitol on seeds, immature embryos, and callus cultures of flax to develop a method for obtaining genotypes resistant to osmotic stress. The work was carried out in the Tver region in the laboratory of breeding technologies in 2017–2019. Flax varieties Barbara, Belinka, LM-98, Aurore, Tverskoy, Svetoch, Diplomat, Symfonia were used as objects of research. The seeds were obtained from the National Flax Collection of the Federal Scientific Center for Bast Crops. The effect of sucrose solution on the length of the primary root was detected at concentrations - 0; 8.7; 14.9%. To assess the germination energy of seeds under osmotic stress, the concentration of sucrose was reduced and the range 0 (control) ... 9% was considered. Immature embryos removed from the capsules on the 10th day after pollination were cultivated on MS medium with sucrose, as a selective agent, at a concentration of 5.0 ... 7.0%. Callus tissues were cultured using mannitol as an osmotic at concentrations of 0; 30.0; 36.4; 37.0; 37.4; 38.0 mg/l. Concentrations of 5.0, 6.0 and 7.0% of sucrose can be used as an osmotic differentiator for seeds (10 ... 80% of seeds germinated in the Belinka variety, 80 ... 100% in the Varbara variety, 80 ... 90% in the variety LM-98). Sucrose, as a selective agent, in a culture of immature flax embryos in vitro at a concentration of 5.0 ... 7.0% can be selective only for certain genotypes, for example, the Aurore variety. The selection of resistant callus cells, followed by the formation of adventive buds and shoots in the meristematic foci, can be carried out on media containing 30.0 or 36.4 mg / L of osmosis, which allows obtaining morphogenic callus, buds, shoots in all studied genotypes, as well as in the Aurore variety 1.1 ... 1.2 byp./callus, in the Tverskoy variety - 0.6 ... 0.8, in the Barbara variety - 1.0 ...1.1

scholarly journals Reaction of flax seeds and explants to osmotic stress

2021 ◽  
Vol 205 (02) ◽  
pp. 56-64
Author(s):  
Elena Vinogradova
Keyword(s):  
Osmotic Stress ◽  
Sucrose Solution ◽  
Primary Root ◽  
Stress Factors ◽  
Federal State ◽  
Scientific Center ◽  
Subsequent Formation ◽  
Fiber Flax ◽  
Callus Tissues

Abstract. Purpose. Obtaining experimental data for the development of methods for obtaining fiber flax forms resistant to stress factors (drought) by cell selection methods. Methods. The work was carried out in the Tver region in the laboratory of breeding technologies in 2017–2019. The flax varieties Varbara, Belinka, LM-98, Svetoch, Diplomat, Symfonia were used as objects of research. The seeds were obtained from the National Flax Collection Federal state budgetary scientific institution “Federal scientific center of lubya kultur”. The effect of sucrose solution on the length of the primary root was detected at concentrations of 0, 8.7, 14.9 %. To assess the energy of seed germination under osmotic stress, the concentration of sucrose was reduced and the range from 0 (control) to 9 % was considered. Results. Sucrose as a selective agent in the culture of immature flax embryos in vitro at a concentration of 5.0–7.0 % can be selective only for certain genotypes, for example, the Svetoch variety. When culturing callus tissues, using mannitol as an osmotic at concentrations of 0; 30.0; 36.4; 37.0; 37.4; 38.0 mg/l, selection of resistant callus cells with subsequent formation in meristematic foci of adventive buds and shoots can be carried out on media containing 30.0 or 36.4 mg/l osmotic. The scientific novelty of the research lies in the fact that the method for obtaining in vitro resistant explants to osmotic stress for flax is being developed for the first time.

scholarly journals USAGE OF CULTURAL FILTRATES IN IN VITRO SELECTION FOR ANTHRACNOSE RESISTANCE

2021 ◽  
Vol 212 ◽  
pp. 92-97
Author(s):  
N. V. Proletova ◽  
Keyword(s):  
In Vitro Selection ◽  
Selective Medium ◽  
Federal State ◽  
Scientific Center ◽  
Morphogenic Callus ◽  
Weakly Virulent ◽  
Vitro Development ◽  
Highly Virulent ◽  
Germinated Seeds

The research was carried out on the basis of the laboratory of selection technologies of the Federal State Budgetary Scientific Institution “Federal Scientific Center of Fiber Crops” (Tver region) in 2018–2020. The aim of the research is in vitro development of new flax genotypes resistant to anthracnose, one of the most harmful fungal diseases. As a result of the research, the composition of the cultural filtrate of the anthracnose causative agent was clarified. It was revealed that toxicity of the cultural filtrates did not depend on the virulence of the strains used in the present studies, the cultural filtrates of strains 784 (highly virulent) and 780 (medium virulent) turned out to be more toxic (decay and death of radicle was observed on the 5th day in 67 - 88% of germinated seeds), less toxic are strains 793 (highly virulent) and 788 (weakly virulent) (decay and death of radicle was observed on the 5th day in 9-15% of germinated seeds). It was found that morphogenic foci were formed more actively in genotypes the morphogenic callus of which was transferred to a medium with a higher concentration of the cultural filtrate; it was shown that in the second passage, when transferring morphogenic calli from a selective medium, which contains 40 ml / L of cultural filtrate on a selective medium also containing 40 ml / L of cultural filtrate, as well as on a selective medium containing 44 ml / L of cultural filtrate, the number of formed morphogenic calli and green buds on the 14th day is significantly higher than in case of transferring on a selective medium containing 36 ml / L of cultural filtrate. Viable regenerant plants were obtained and genotypes were isolated, which retained resistance to anthracnose for three generations at a level of 50 - 60%: NO-78 x Lenok, NL-103-2 x Lenok, NL-40-1 x Lenok, NE-38 x Rosinka, NE-36 x Lenok, NE-17 x Lenok, NE-16-2 x Rosinka.

BIOTECHNOLOGICAL METHODS - A TOOL FOR CREATING NEW FLAX GENOTYPES RESISTANT TO ANTHRACNOSE

2020 ◽  
Keyword(s):  
Culture Filtrate ◽  
Embryo Culture ◽  
Selective Medium ◽  
Field Resistance ◽  
Immature Embryos ◽  
Cell Selection ◽  
Morphogenic Callus ◽  
Morphogenetic Potential ◽  
Regenerant Plants

The purpose of this research was to create new flax genotypes resistant to anthracnose using biotechnological techniques and methods. As a result of studies with using a culture of immature embryos and a selective medium, flax regenerant plants resistant to the culture filtrate (CF) of the fungus - anthracnose pathogen Colletotrichum lini Manns et Bolley and line 21 resistant to this pathogen were obtained. A scheme for differentiating flax genotypes in vitro by resistance to anthracnose has been developed. It was established that upon cultivation of immature embryos on Sh-2 medium containing CF of the fungus, the causative agent of anthracnose, of a mixture of strains 680, 677 * 674, 674 * at a concentration of 36.0 ml / l, the amount of morphogenic callus formed in the first and second passages, expressed in percent, and the indicator of the field resistance of this genotype to anthracnosis on an artificial infectious-provocative background was close in value, and the number of morphogenic callus formed in the first and second passages can be used to judge the resistance of the studied genotypes to ant cancer and differentiate them by resistance to this pathogen. The influence of the flax genotype on the potency of cells to morphogenesis under selective conditions was revealed. Cells of genotypes L 957-8-7, Alexim, Punjab, Zaryanka had high morphogenetic activity. The morphogenetic potential of genotypes L 1506-8-4, Rosinka has already been exhausted by 2 ... 3 passages. It was found that biotechnological methods: cell selection in vitro, embryo culture are effective in creating genotypes of flax, more resistant to anthracnose than original forms.

scholarly journals Creation of clary sage cultivar using cell engineering methods. 1. Obtaining of plant-regenerants in callus culture in vitro

2021 ◽  
Vol 1 (25) ◽  
pp. 98-112
Author(s):  
N.A. Yegorova ◽  
◽  
I.V. Stavtzeva ◽  
Keyword(s):  
Callus Formation ◽  
Leaf Shape ◽  
Flower Color ◽  
Callus Cultures ◽  
Maximum Frequency ◽  
Agricultural Plant ◽  
Morphogenic Callus ◽  
Somaclonal Variability ◽  
Callus Tissue Culture

To increase the efficiency in agricultural plant breeding, including clary sage – one of the main essential oil crops grown in Russia, it is necessary to use biotechnological methods. One of these techniques is based on the induction of somaclonal variability in the callus tissue culture. To develop it, it is necessary to optimize the conditions for obtaining plant-regenerants in vitro and their analysis. The aim of this work was to study the features of morphogenesis and regeneration of plants from callus cultures to develop cell technologies for creating an initial breeding material based on somaclonal variability in Salvia sclarea L. In the course of the research, we found that the optimal explants for obtaining morphogenic callus, from which shoots were regenerated, were segments of buds and stems with a node (isolated from seedlings in vitro). Cytological analysis of callus cultures revealed two types of morphogenesis – organogenesis (gemmogenesis) and somatic embryogenesis. The features of the morphogenic callus formation of six sage cultivars and samples during the long-term cultivation were studied. The maximum frequency of morphogenesis was noted in the 2nd passage (from 32.4 to 85.2 %, depending on the genotype). Then, to the 8–10th passage, this indicator decreased to 0.0–3.9 %.‘S-785’ and ‘Taigan’ cultivars showed the highest morphogenesis frequency (81.5–85.2 %) and duration of callus regeneration potential (up to the 10th passage). The analysis of callus cultures of six donor plants of ‘S-785’ cultivar helped us to reveal their heterogeneity in morphogenesis induction ability. The maximum frequency of morphogenic callus formation (76.3–91.5 % in the 2nd passage) and the duration of the morphogenic potential preservation (up to the 12th passage) were observed in plants No. 3 and 9, whereas in No. 2, regeneration with a frequency of 3.6–9.7 % was observed only during three passages. Analysis of plants obtained from calli showed their variability in morphology – up to 12.5 % of the samples had deviations compared to the initial cultivar ‘S-785’ in leaf shape, inflorescence structure, flower color, etc. Somaclonal changes in morphological and economically useful traits revealed in regenerants indicate that they are promising for use in sage breeding.

scholarly journals Morphogenic processes in callus tissue cultures and de novo regeneration of plants in Actinidia chinensis Planch

2014 ◽  
Vol 67 (3-4) ◽  
pp. 217-222 ◽  
Author(s):  
Adela Ludvová ◽  
Mária G. Ostrolucká
Keyword(s):  
De Novo ◽  
Leaf Explants ◽  
Structural Heterogeneity ◽  
Callus Cultures ◽  
Culture Conditions ◽  
Actinidia Chinensis ◽  
Indirect Organogenesis ◽  
Vascular Elements ◽  
Callus Tissues

Our experiments have confirmed the considerable disposition of leaf explants of <em>Actinidia chinensis</em> Planch. for induction and intensive proliferation of callus cultures, as well as, a possibility to regulate morhogenesis in in vitro conditions. Under specific culture conditions the morphogenic potential of callus cells of <em>Actinidia chinensis</em> was manifested both in organogenesis and somatic embryogenesis. Organogenesis was represented by induction of adventitious buds and regeneration shoots on the modified MS culture medium (Murashige and Skoog 1962) with BAP in combination with GA<sub>3</sub> (each 1.0 mg. l<sup>-1</sup>). Rooting of shoots was successful on modified MS medium containing IBA (0.5-1.0 mg. l<sup>-1</sup>). Histological studies of callus tissues revealed their structural heterogeneity. Morphogenic processes in the callus were characterized by the appearance of meristematic zones and vascular elements. The formation of apical meristem, leaf primordia and finally shoot development proved de novo regeneration in callus culture. The obtained results demonstrate a possibility of plant regeneration through indirect organogenesis, which can be used for propagation of<em> Actinidia chinensis</em> Planch.

scholarly journals Fungal Colonization and Host Defense Reactions in Ulmus americana Callus Cultures Inoculated with Ophiostoma novo-ulmi

2009 ◽  
Vol 99 (6) ◽  
pp. 642-650 ◽  
Author(s):  
Mirella Aoun ◽  
Danny Rioux ◽  
Marie Simard ◽  
Louis Bernier
Keyword(s):  
Electron Microscopy ◽  
Quantitative Polymerase Chain Reaction ◽  
Callus Cultures ◽  
Dutch Elm Disease ◽  
Fungal Colonization ◽  
Ulmus Americana ◽  
In Vitro System ◽  
Transmission Electron ◽  
Callus Tissues

The host–pathogen interaction leading to Dutch elm disease was analyzed using histo- and cyto-chemical tests in an in vitro system. Friable and hard susceptible Ulmus americana callus cultures were inoculated with the highly aggressive pathogen Ophiostoma novo-ulmi. Inoculated callus tissues were compared with water-treated callus tissues and studied with light microscopy (LM), transmission-electron microscopy (TEM), and scanning-electron microscopy (SEM). New aspects of this interaction are described. These include the histological observation, for the first time in plant callus cultures, of suberin with its typical lamellar structure in TEM and the intracellular presence of O. novo-ulmi. Expression of the phenylalanine ammonia lyase gene, monitored by real-time quantitative polymerase chain reaction, was correlated with the accumulation of suberin, phenols, and lignin in infected callus cultures. This study validates the potential use of the in vitro system for genomic analyses aimed at identifying genes expressed during the interaction in the Dutch elm disease pathosystem.

scholarly journals Multi-Step Cellular and Tissue Selection of Soybean for Resistance to Osmotic Stress Using Peg 6000 Invitro Conditions

2021 ◽  
Vol 8 (4) ◽  
Author(s):  
Mohammad Naim Noori
Keyword(s):  
Polyethylene Glycol ◽  
Osmotic Stress ◽  
Nutrient Medium ◽  
Selective Agent ◽  
Soybean Variety ◽  
Peg 6000 ◽  
Final Volume ◽  
Polyethylene Glycol 6000 ◽  
Selection Of

Cell and tissue selection in vitro allows targeted selection of the desired traits under severe selective conditions at the level of individual cells and tissues. On the basis of multistage cell and tissue selection with the use of a selective agent - neutral osmotic polyethylene glycol 6000 in increasing concentrations -5%, 10%, 15%, 20% of the final volume of the nutrient medium, callus of the Zhansaya soybean variety that are stably resistant to osmotic stress have been obtained. Regeneration from callus was noted only in8.3 % of the planted callus 6 soybean regenerants plants resistant to osmotic stress were obtained from callus.

scholarly journals Glyphosate selection of maize transformants containing CP4epsps gene

2020 ◽  
Vol 26 ◽  
pp. 239-244
Author(s):  
I. O. Nitovska ◽  
B. V. Morgun ◽  
O. Ye. Abraimova ◽  
T. M. Satarova
Keyword(s):  
Transgenic Plants ◽  
Dna Analysis ◽  
Selective Agent ◽  
Regeneration Rate ◽  
Agrobacterium Mediated Transformation ◽  
Plant Dna ◽  
Morphogenic Callus ◽  
Selection Of ◽  
Maize Callus

Aim. To study the selection conditions of maize transformants containing the CP4epsps gene using glyphosate as a selective agent. Methods. Tissue culture in vitro, Agrobacterium-mediated transformation, selection of transgenic plants, isolation of total plant DNA, analysis of plant DNA by polymerase chain reaction (PCR). Results. The morphogenic maize callus of immature embryos of the hybrid (PLS61)R2×PLS61 was produced, which had a high regeneration rate (up to 95%), that persisted over long cultivation. Agrobacterium mediated transformation of the morphogenic callus and selection of the transgenic material using glyphosate yielded maize transformants containing the CP4epsps gene at a frequency of 1%. Conclusions. Maize genotype (PLS61)R2×PLS61 is promising for studies on the maize genetic transformation, in particular for the production of transgenic maize resistant to glyphosate herbicide. The use of morphogenic maize callus (PLS61)R2×PLS61 and glyphosate as a selective agent at a concentration of 0.1 mM and 0.25 mM in media for callusogenesis and 0.01 mM in the medium for regeneration was effective for the selection of transgenic plants with the gene CP4epsps. Keywords: Zea mays L., morphogenic callus, Agrobacterium-mediated transformation, PCR, genetic engineering.

scholarly journals ВМІСТ ФЛАВОНОЇДІВ І КCАНТОНІВ У КАЛЮСНИХ КУЛЬТУРАХ РОСЛИН ВИДІВ РОДУ GENTIANA L. ЗА ВИРОЩУВАННЯ У РІДКОМУ ЖИВИЛЬНОМУ СЕРЕДОВИЩІ НА ПОРОЛОНОВИХ ПІДКЛАДКАХ

2020 ◽  
Vol 80 (3-4) ◽  
pp. 39-49
Author(s):  
L. R. Hrytsak ◽  
V. M. Mel’nyk ◽  
M. Z. Prokopyak ◽  
O. Yu Mayorova ◽  
Kh. M. Kolisnyk ◽  
...  
Keyword(s):  
Nature Reserve ◽  
Agar Medium ◽  
Growth Index ◽  
Callus Cultures ◽  
Biologically Active ◽  
Wild Plants ◽  
Growth Media ◽  
Nutrient Media ◽  
Callus Tissues

The content of flavonoids and xanthones in callus cultures derived from the roots of plants of six species of Gentiana L. genus was studied during the cultivation of these cultures in liquid growth media on foam substrates. The research findings indicate that for most callus cultures, which were grown on both agar and foam substrates, the content of biologically active substances (BAS) was higher or close to that in the roots of wild plants, but lower compared to their shoots. The content of flavonoids and xanthones in tissue cultures grown in liquid nutrient media exceeded (G. punctata, Mt. Breskul, G. asclepiadea, Mt. Pozhyzhevska, G. cruciata, Krenychi village and G. lutea, Mt. valley Rohnechska), was close (G acaulis, Mt. Turkul) or lower (G. cruciata, «Medobory» Nature Reserve, G. lutea, Mt. Troyaska) compared to those in the corresponding calluses on agar substrates. In the callus of G. pneumonanthe (Vyhoda village) during cultivation on agar medium and in liquid medium on foam substrates, flavonoids and xanthones were not detected. For most callus cultures of gentians: G. punctata (Mt. Breskul), G. asclepiadea (Mt. Pozhyzhevska), G. cruciata (Krenychi village), G. lutea (Mt. valley Rohnechska), cultivation in a liquid growth medium on foam substrates can increase the growth of callus biomass (1.3–1.7 times) and the content of flavonoids (1.2–1.6 times) and xanthones (1.2–2.3 times) in comparison with the same cultures on agar media. For the callus of G. cruciata («Medobory» Nature Reserve) and G. lutea (population of Mt. Troyaska) on liquid media with foam substrates, both the growth index by fresh weight and the content of secondary metabolites are lower compared to cultures from agar medium. The growth of callus G. acaulis (Mt. Turkul) on the nutrient medium with foam substrates is more intense than on agar, but with lowered BAS. Thus, the developed method of cultivating callus tissues of gentians in liquid nutrient media on foam substrates can reduce costs by replacing agar with foam substrates, as well as increase both the yield of biomass of most callus cultures and their ability to synthesize and accumulate flavonoids and xanthones. Key words: Gentiana L. species, in vitro cultures, foam substrates, flavonoids, xanthones.

scholarly journals In Vitro Callus Culture of Dianthus Chinensis L. for Assessment of Flavonoid Related Gene Expression Profile

2021 ◽  
Author(s):  
R. Sreelek ◽  
Elenjikkal A Siril
Keyword(s):  
Gene Expression ◽  
Gene Expression Profile ◽  
Expression Profile ◽  
Light Condition ◽  
Callus Cultures ◽  
Biological Research ◽  
Dark Condition ◽  
Related Gene ◽  
Callus Tissues

Abstract Dianthus chinensis L. is an edible, ornamental herb used to prepare the Dianthi Herba, a Chinese traditional rejuvenating medicine. Owing to the rapid proliferation of callus tissues, in vitro production of flavonoids has their own specific importance. Callus cultures raised followed by auxin directed biosynthesis of flavonoid through related transcript profile were carried out. Murashige and Skoog (MS) medium fortified with 2,4- Dichlorophenoxy acetic acid (2,4- D) or picloram induced formation of friable callus from internode derived cultures of D. chinensis. Culture medium containing 2,4- D (10 µM) produced the highest flavonoid content, 4.44 mg quercetin equivalent per gram (QE g− 1) under incubation in continuous dark condition, while maximum dry weight yield (0.38 g/ culture) was obtained from 10 µM 2,4- D under 16 h light / 8 h dark condition (50 µmol m− 2 s− 1 irradiance) at 60 days of incubation. The callus raised in light condition in 10 µM 2,4- D selected to analyze flavonoid related gene expression profile viz., chalcone synthase (CHS), chalcone isomerase (CHI), flavanone-3-hydroxylase (F3H), and flavonol synthase (FLS) at specific time intervals. The transcript abundance of CHS, F3H, or FLS gene was higher at 60 days old callus cultures and reaching 11.59, 48.31, and 114.63-fold relative expression than that of initial callus tissues respectively. These understandings are critical for the regulation of targeted phytochemicals as well as their wide exploitation in the field of biological research.

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